Real-Time Temperature Sensing Using a Ratiometric Dual Fluorescent Protein Biosensor

Sorenson, Alanna E., and Schaeffer, Patrick M. (2023) Real-Time Temperature Sensing Using a Ratiometric Dual Fluorescent Protein Biosensor. Biosensors, 13 (3). 338.

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Accurate temperature control within biological and chemical reaction samples and instrument calibration are essential to the diagnostic, pharmaceutical and chemical industries. This is particularly challenging for microlitre-scale reactions typically used in real-time PCR applications and differential scanning fluorometry. Here, we describe the development of a simple, inexpensive ratiometric dual fluorescent protein temperature biosensor (DFPTB). A combination of cycle three green fluorescent protein and a monomeric red fluorescent protein enabled the quantification of relative temperature changes and the identification of temperature discrepancies across a wide temperature range of 4–70 °C. The maximal sensitivity of 6.7% °C−1 and precision of 0.1 °C were achieved in a biologically relevant temperature range of 25–42 °C in standard phosphate-buffered saline conditions at a pH of 7.2. Good temperature sensitivity was achieved in a variety of biological buffers and pH ranging from 4.8 to 9.1. The DFPTB can be used in either purified or mixed bacteria-encapsulated formats, paving the way for in vitro and in vivo applications for topologically precise temperature measurements.

Item ID: 78193
Item Type: Article (Research - C1)
ISSN: 2079-6374
Keywords: GFP; mCherry; ratiometric; DSF-GTP; fluorescent; thermal; temperature; differential scanning fluorimetry; DFPTB
Copyright Information: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (
Date Deposited: 03 Aug 2023 03:42
FoR Codes: 31 BIOLOGICAL SCIENCES > 3106 Industrial biotechnology > 310604 Industrial biotechnology diagnostics (incl. biosensors) @ 100%
SEO Codes: 28 EXPANDING KNOWLEDGE > 2801 Expanding knowledge > 280102 Expanding knowledge in the biological sciences @ 100%
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